Plant Tissue Fructose (FT) Content Assay Kit (Resorcinol, Micro Method)

Cat. No.: P1508220

GRADE & PURITY BioReagent

Overview

Fructose is one of the most common ketohexoses and an isomer of glucose. It exists in large amounts in the free state in fruit juices and honey and can combine with glucose to form sucrose. Fructose is the sweetest monosaccharide and is widely used in the production of food, pharmaceuticals, and health products.

Detection Principle

Under acidic conditions, fructose reacts with resorcinol to produce a colored substance with a characteristic absorption peak at 480 nm.

Note: Before formal testing, it is recommended to perform a preliminary experiment with 2-3 samples expected to show significant differences.

Reagents, consumables and Equipments not provided

• Microplate reader or visible spectrophotometer (capable of measuring absorbance at 480 nm)

• 96-well plate or micro-volume glass cuvette, Adjustable pipettes and tips

• Constant temperature water bath, Centrifuge, Analytical balance

• Deionized water

• Homogenizer

Procedure

1. Reagent Preparation

Reagent Name	Preparation	Notes
Extraction Buffer	Ready-to-use. Equilibrate to room temperature before use.	Store at 4°C.
Working Reagent Ⅰ	Prepare immediately before use. Dissolve Reagent Ⅰ in 1 mL deionized water. Take 100 µL of this solution and add to 900 µL deionized water. Mix thoroughly.	Unused reagent can be stored at 4°C protected from light for up to two weeks.
Reagent Ⅱ	Ready-to-use. Equilibrate to room temperature before use.	Store at 4°C.
Reagent Ⅲ	Ready-to-use. Equilibrate to room temperature before use.	Store at 4°C protected from light.
Reagent Ⅳ	Ready-to-use.	Store at room temperature protected from light.

Note: Reagent Ⅱ and Reagent Ⅲ are toxic and have an irritating odor. It is recommended to perform the experiment in a fume hood.

2. Sample Preparation

*Note: Fresh samples are recommended. If not used immediately, samples can be stored at -80°C for several weeks. Control the thawing temperature and time during measurement. When thawing at room temperature, complete the process within 4 hours.*

Weigh 0.1-0.2 g of sample and grind at room temperature. Add 0.5 mL of Extraction Buffer, grind appropriately, and quickly transfer to a capped centrifuge tube. Place in an 80°C water bath for 10 minutes (keep cap tight to prevent water loss), vortex 3-5 times. After cooling, centrifuge at 4,000 g, 25°C for 10 minutes. Collect the supernatant. Add a small amount (approx. 2 mg) of Reagent Ⅳ. Decolorize at 80°C for 30 minutes (keep cap tight). Add another 0.5 mL of Extraction Buffer, centrifuge at 4,000 g, 25°C for 10 minutes. Collect the supernatant for testing.

3. Experimental Steps

3.1 Preheat the microplate reader or visible spectrophotometer for 30 minutes and set the wavelength to 480 nm. Zero the visible spectrophotometer with deionized water.

3.2 Operation Table (perform in 1.5 mL EP tubes):

Reagent (µL)	Test Tube	Blank Tube	Standard Tube
Sample Supernatant	30	0	0
Working Reagent Ⅰ	0	0	30
Deionized Water	0	30	0
Reagent Ⅱ	210	210	210
Reagent Ⅲ	60	60	60

3.3 Mix well. React in a 95°C water bath for 30 minutes (keep cap tight to prevent water loss). After cooling, transfer 200 µL to a micro-volume glass cuvette or 96-well plate. Measure the absorbance at 480 nm, recorded as A_Test, A_Blank, A_Standard. Calculate ΔA_Test = A_Test - A_Blank and ΔA_Standard = A_Standard - A_Blank.

*Note: The blank and standard tubes only need to be prepared 1-2 times. It is recommended to perform a preliminary experiment with 2-3 samples showing expected large differences before formal testing. If A_Test is less than 0.05, appropriately increase the sample amount. If A_Test is greater than 2.0, further dilute the sample with Extraction Buffer and multiply the result by the dilution factor, or reduce the amount of sample used for extraction.*

4. Calculation of Results

Note: We provide two formula sets: the derived calculation formulas and the simplified formulas. They are completely equivalent. The simplified formulas are recommended as the final calculation formulas.

Fructose Content Calculation:

Fructose Content (mg/g fresh weight) = (C_Standard × V_Sample) × ΔA_Test ÷ ΔA_Standard ÷ (W × V_Sample ÷ V_{Total Extract}) = ΔA_Test ÷ ΔA_Standard ÷ W

Parameter Description:

C_Standard: Standard tube concentration, 1 mg/mL;

V_Sample: Volume of sample added, 0.03 mL;

V_{Total Extract}: Volume of extraction buffer added, 1 mL;

W: Sample fresh weight, g.

5. Example Results

Figure 1. Fructose content in melon and orange determined by this kit

Notes

This product is for scientific research use only and is not intended for clinical diagnosis. For your safety and health, please wear a lab coat and disposable gloves during operation.

Storage and Shipping

Storage

Store at 2-8°C,Protected from light,Room temperature

Shipped In

Wet ice

Stability And Storage

Each component has a shelf life of 1 year under corresponding storage conditions. P1508220A, P1508220C: Store at 2-8℃ long term (12 months). P1508220B, P1508220D: Store at 2-8℃ long term (12 months). Store in the dark. P1508220E: Store at room temperature

Contents & Storage

P1508220	Component	48T	96T	Storage
P1508220A	Extraction Buffer	60 mL	120 mL	2-8℃
P1508220B	Reagent Ⅰ	1EA	1EA	2-8℃. Store in the dark.
P1508220C	Reagent Ⅱ	13 mL	26 mL	2-8℃
P1508220D	Reagent Ⅲ	4 mL	8 mL	2-8℃. Store in the dark.
P1508220E	Reagent Ⅳ	1EA	1EA	RT. Store in the dark.